Mono, Di &amp; Tri Cyclic Compounds Useful for Lowering IgE Concentrations

ABSTRACT

The present invention is directed to novel compounds which are derivatives of minocycline or doxycycline, pharmaceutical compositions containing same and use thereof in lowering excess IgE levels in a mammal suffering from a disease where IgE is pathogenic.

BACKGROUND OF THE INVENTION

This invention relates to novel derivatives of minocycline anddoxycline, pharmaceutical compositions containing same and their use inlowering IgE levels in mammals, especially humans, suffering from adisease where IgE is pathogenic, such as allergies, asthma, especiallyhuman allergic response, and diseases associated with an inflammatoryresponse.

Diseases involving inflammation are characterized by the influx ofcertain cell types and mediators, the presence of which can lead totissue damage and sometimes death. Diseases involving inflammation areparticularly harmful when they afflict the respiratory system, resultingin obstructed breathing, hypoxemia, hypercapnia and lung tissue damage.Obstructive diseases of the airways are characterized by air flowlimitation (i.e., airflow obstruction or narrowing) due to constrictionof airway smooth muscle, edema and hypersecretion of mucous leading toincreased work in breathing, dyspnea, hypoxemia and hypercapnia.

A variety of inflammatory agents can provide air flow limitation, suchas for example, allergens. In particular, allergens and other agents inallergic or sensitized animals (i.e., antigens and haptens) cause therelease of inflammatory mediators that recruit cells involved ininflammation. Such cells include lymphocytes, eosinophils, mast cells,basophils, neutrophils, macrophages, monocytes, fibroblasts andplatelets. A variety of studies have linked the degree, severity andtiming of the inflammatory process with the degree of airwayhyperresponsiveness. Thus, a common consequence of inflammation isairflow limitation and/or airway hyperresponsiveness.

Asthma is a significant disease of the lung which is typicallycharacterized by periodic air flow limitation and/or hyperresponsivenessto various stimuli which results in excessive airways narrowing. Othercharacteristics can include inflammation of airways and eosinophila.More particularly, allergic asthma is often characterized byeosinophilic airway inflammation and airway responsiveness.

An estimated 16 million persons in the U.S. have asthma, which is about10% of the population. The numbers have increased about 25% in the last20 years. The estimated cost of treating asthma in the U.S. exceeds $6billion. About 25% of patients with asthma who seek emergency carerequire hospitalization. The largest single direct medical expenditurefor asthma has been in patient hospital services (emergency care), at acost of greater than $1.6 billion. The cost for prescription medicationsis at least $1.1 billion.

According to the National Ambulatory Medical Care Survey, asthmaaccounts for 1% of all ambulatory care visits and the disease continuesto be a significant cause of missed school days in children. Despiteimproved understanding of the disease process and better drugs, asthmamorbidity and mortality continues to rise in this country and worldwide.Thus, asthma constitutes a significant public health problem.

The pathophysiologic processes that attend the onset of an asthmaticepisode can be broken down into essentially two phases, both marked bybronchioconstriction, that causes wheezing, chest tightness, anddyspnea. The first, early phase asthmatic response is triggered byallergens and irritants. Allergens cross-link immunoglobulin (IgE)molecules bound to receptors on mast cells and basophils, causing themto release a number of pre-formed inflammatory mediators, includinghistamine. Additional triggers include the osmotic changes in airwaytissues following exercise and/or the inhalation of cold, dry air. Thesecond, late phase response that follows is characterized byinfiltration of activated eosinophilis and other inflammatory cells intoairway tissues, epithelial desquamation and by the presence of highlyviscous mucus within the airway. The damage caused by this inflammatoryresponse leaves the airways “primed” or sensitized, such that smallertriggers are required to elicit subsequent asthma symptoms.

For instance, human allergic asthma, a disease characterized by airwayhyperresponsiveness and bronchial inflammation, is mediated by a varietyof activated leukocytes, including eosinophils, mast cells, CD4+ Tlymphocytes, and CD19+ B cells.

Current treatments, which improve airway hyperresponsivenss, includevarious anti-inflammatory agents, which reduce mucosal inflammation andasthma pathogenesis; however their efficacies vary markedly.

Short acting β₂-adrenegric agonists, terbutaline and albuterol, long themainstay of asthma treatment, act primarily during the early phase asbronchodilators. The newer long acting β₂ agonists do not possesssignificant anti-inflammatory activity; they have no effect on bronchialhyperreactivity.

Numerous other drugs target specific aspects of the early or lateasthmatic responses. For example, antihistamines, like loratadine,inhibit early histamine-mediated inflammatory responses. Otherantihistamines, such as azelastine and ketotifen, have bornanti-inflammatory and weak bronchodilatory effects, but they currentlydo not have any established efficacy in asthma treatment.

Phosphodiesterase inhibitors, like theophylline/xanthines, may attenuatelate inflammatory responses, but there is no evidence that the compoundsdecrease bronchial hyperreactivity. Anticholinergics, like ipratropiumbromide, which are used in cases of acute asthma to inhibit severebronchoconstriction, have no effect on early or late phase inflammation,no effect on bronchial hyperreactivity and therefore essentially have norole in chronic therapy.

The corticosteroid drugs, like budesonide, are among the most potentanti-inflammatory agents. Inflammatory mediators or release inhibitors,like cromolyn and nedocromil, act by stabilizing mast cells andinhibiting the late phase inflammatory response to allergen. Thus,cromolyn and nedocromil, as well as the corticosteroids, all reducebronchial hyperactivity by minimizing the sensitivity effect ofinflammatory damage to the airways. These anti-inflammatory agents,however, do not produce bronchodilation.

Thus, while numerous drugs are currently available for the treatment ofasthma, these compounds are primarily palliative and/or have significantside effects.

Unfortunately, none of the aforementioned drugs target the underlyingcause of asthma.

Consequently, new therapeutic approaches which target the underlyingcause rather than the cascade of symptoms would be highly desirable. Thepresent inventors have searched for the underlying cause of asthmas,especially human allergic asthma. In asthma, CD4+ T cells secrete IL-4,a (Th-)2 type cytokine, which is required for IgE production and whichis implicated in airway hyperresponsiveness, as well other cyclokineswhich increase IgE production.

The present inventors have found that a tetracycline congener,minocycline or doxycycline suppress human and murine IgE production.More specifically, it has been found that minocycline and doxycyclinelower IgE concentrations. Further, it has been found that minocyclinesuppresses human and murine IgE production in vivo and that minocyclineand another tetracycline congener, doxycycline, suppresses human IgEproduction in vitro.

It has been reported that allergic steroid dependent asthmatic patientstreated with an oral administration of minocycline improved theirsymptoms (A.M. and P.M.), and decreased oral corticosteroid requirementsSee, Joks, et al., J. Allergy Clin. Immunol. 1998, 101:562. Additionalstudies of O'Dell, et al. in Arthritis Rheum: 1997, 40:842-848 andArthritis Rheum: 1999, 42:1691-1695 have shown that treatment of mildand moderate rheumatoid arthritis (RA) patients with minocycline had noside effects, and appears to be an effective therapy for early RA.Moreover, Yu, et al. in Arthritis Rheum: 1992, 35:1150-1155 reportedthat treatment of dogs with minocycline or doxycycline reduced theseverity of osteoarthritis (OA), while studies by Thong, et al. in ClinExp Immunol., 1979, 35:443-446, have shown that doxycycline andtetracycline inhibit the ability of mice to mount delayed-typehypersensitivity responses. Further, studies in vitro have demonstratedthat treatment of human whole blood cultures with minocycline ortetracycline at physiological doses inhibits mitotic responses tophytohemagglutinin (See Ingham, et al., Antimicrob Chemother. 1991,27:607-617) and inhibits inducible nitric oxide synthase (iNOS)expression by murine macrophages See Amin, et al., PNAS, 1996,93:14014-14019. In addition, it has been found that both minocycline anddoxycycline suppress anti-CD40 rhIL-4 mediated in vitro induction of IgEresponses by patient PBMC (peripheral blood mononuclear cells) in a dosedependent manner. (Smith-Norowitz, et al. Annals of Allergy, Asthma &,Immunology, 2002, 89:8, 172-179. In addition, it has been shown thatdoxycycline suppresses PHA/IL-4 mediated IgE response by normal mousespleen cells. Kuzin, et al. International Immunol 12, 921-931 (2000).

Although minocycline and doxycycline show effectiveness in suppressingexcess IgE levels, being tetracyclines, they have the same adverseeffects associated with tetracyclines. For example, a high percentage ofpatients are unable to tolerate oral tetracyclines for extendedperiod's. The intolerance to tetracyclines can manifest itself intogastrointestinal problems, e.g., epigastric pain, nausea, vomiting anddiarrhea or other problems related to taking the tetracyclines for longterm treatment, such as mucosal candidiasis, staining teeth and thelike.

The objective of the present invention is to find molecules thatsuppress IgE products, and reduce and/or eliminate the aforementionedside effects associated with tetracyclines. The present invention isdirected to a means of achieving this objective by making modificationsof the structures of the minocycline and doxycycline. More specifically,the present invention is directed to these new molecules and their usein suppressing excess IgE levels in patients suffering from asthma,allergies, inflammatory conditions, or other diseases where IgE ispathogenic.

SUMMARY OF THE INVENTION

The present invention is directed to the following compounds which areeffective in lowering excess IgE levels in plasma:

and to the pharmaceutically effective salts thereof.

The present invention is also directed to pharmaceutical compositionscomprising a therapeutically effective amount of a compound selectedfrom the group consisting of Formulae I, II, III, IV, V, VI, VII, andVIII or pharmaceutically acceptable salts thereof and a pharmaceuticalcarrier thereof. In addition, the present invention is directed to amethod of lowering excess IgE concentration in the plasma of a mammalcomprising administering thereto a compound of Formulae I-VIII orpharmaceutically acceptable salts thereof. A further embodiment isdirected to treating a mammal suffering from a disease where IgE ispathogenic which method comprises administering to a mammal sufferingtherefrom an IgE lowering effective amount of a compound selected fromthe group consisting of Formulae I, II, III, IV, V, VI, VII and VIII orpharmaceutically acceptable salts thereof or combination thereof.Diseases in which the IgE is pathogenic include allergies and asthma,including human allergic asthma and inflamatory condition. In thesediseases, the IgE concentration in the plasma is higher than normal. Afurther embodiment is directed to the prophylaxis of a mammal having anexcess level of IgE in the plasma from suffering from a disease k whichIgE is pathogenic which comprises administering a prophylacticallyeffective amount of a compound of Formulae I-VIII or pharmaceuticallyacceptable salts thereof.

DETAILED DESCRIPTION

Each of the compounds of Formulae I-VIII and pharmaceutically acceptablesalts thereof are derivatives of minocycline and doxycline. Thestructures of minocycline and doxycline are as follows:

The inventors subjected these molecules to in silico tools tocomputationally analyze their molecular shapes, and as a result thereofthey identified regions of overlap, which may correspond to the activesites of the molecules responsible or attributable for IgE suppression.They noted that the lower portions of the parent molecules are rich inketo-enol capabilities. Based on these calculations, they deduced thecompound of Formula I, which can effect the lowering of excess IgE inthe plasma.

In addition, they noted that the A ring of these tetracyclines containeda ketone, dimethyl amino and an amide group. Thus, they deduced that astructure containing just the A ring of Formula II would also lowerexcess IgE in the plasma. Moreover, they also deduced the structures ofIII and IV which are bicyclic structures containing the A & B ring ofdoxycycline and minocycline, respectively, having IgE suppressiveactivity. In addition, they deduced structures of Formula V and VI whichcontain the A, B & C rings of doxycycline and minocycline, respectively,having IgE suppressive activity. Finally, they deduced structures VIIand VIII, which contains the A ring without the amide at position 2 orthe hydroxy groups at position 3, respectively, which also has IgEsuppressive activity.

As used herein the term “patient” or “subject” refers to a warm bloodedanimal and preferably mammals, such as for example, cats, dogs, horse,cows, pigs, mice, rats and primate including humans. The preferredpatient is humans.

Unless indicated to the contrary, the term “drug” herein is used toconnote one of the aforementioned a compound which is used to lower theIgE concentration in the plasma of a mammal, e.g., human.

The compounds of the present, invention that are basic in nature(Compounds II-VIII) are capable of forming a wide variety of salts withvarious inorganic and organic acids. The acids that may be used toprepare pharmaceutically acceptable salts.

The term “pharmaceutically acceptable salt” embraces salts commonly usedto form alkali metal salt or for addition salts of free bases containingamine functionalities. The nature of the salt is not critical, providedthat it is pharmaceutically acceptable. Suitable pharmaceuticallyacceptable acid addition salts of compounds containing aminefunctionalities such as present in compounds of Formulae II-VIII may beprepared from an inorganic or organic acids. Examples of such inorganicsalts are hydrochloric, hydrobromic, nitric, carbonic, sulfuric andphosphoric. Appropriate organic acid salts may be selected fromaliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic,carboxylic and sulfonic classes of organic acids, examples of whichinclude but not limited to, formic, acetic, propionic succinic,glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic,glucoronic, maleic, fumaric, benzoic, methanesulfonic, benzenesulfonic,phenylacetic, stearic and the like.

Although such salts must be pharmaceutically acceptable salts foradministration to a subject, e.g., mammal, it is preferably to preparesuch salt from the free base (the free amine) is depicted in compoundsof Formulae II-VIII by treating the same with a substantially equivalentamount of the chosen mineral or organic acid in an aqueous solventmedium or suitable organic solvent, such as methanol or ethanol. Uponevaporation of the solvent, the desired pharmaceutically acceptable saltis obtained.

Thus, as used herein when referring to the expression “one or morepharmaceutically acceptable salts thereof”, it is to be understood thatthe term refers to “pharmaceutically acceptable salts” of compounds ofFormulae II-VIII.

An embodiment of the present invention is directed to the use of IgElowering effective amounts of a compound of Formulae I, II, III, IV, V,VI, VII or VIII or pharmaceutically acceptable salts of compounds ofFormulae II-VIII or combination thereof for the suppression of elevatedconcentrations relative to normal concentrations of IgE in the bloodplasma of a patient suffering from a disease in which IgE is pathogenic.Accordingly, compounds of Formulae I-VIII, singly or in combination, orpharmaceutically acceptable salts of the compounds of Formulae II-VIIIin amounts effective in lowering the concentration of excess IgE in theplasma are useful for treating human allergic asthma. In addition, thesecompounds are anti-inflammatory agents and are useful, either singly orin combination to treat inflammatory conditions when administered topatients in IgE lowering effective amounts, as defined herein.

The compounds, when administered singly, are administered therefore intherapeutically effective amounts. If administered in combination, theyare administered in total in therapeutically effective amounts.

The physician will determine the dosage of the compounds of FormulaeI-VIII or pharmaceutically acceptable salts of compounds of FormulaeII-VIII which will be most suitable and it will vary with the form ofadministration and the particular compound chosen, and furthermore, itwill vary depending upon various factors, including but not limited tothe patient under treatment, the age of the patient, the severity of thecondition being treated and the like. The physician will generally wishto initiate treatment with small dosages substantially less than theoptimum dose of the compound and increase the dosage by small incrementsuntil the optimum effect under the circumstances is reached. Thesecompounds, when given orally, are administered preferably in dosagesranging from about 1 to about 400 mg/day and more preferably from about1 to about 300 mg/day. When given parenterally, the compounds of thepresent invention, are administered preferably in dosages of, forexample, about 1.5 to about 400 mg/day, and more preferably from about 1to about 300 mg/day also depending upon the host and the severity of thecondition being treated and the compound utilized.

This dosage regimen may be adjusted by the physician to provide theoptimum therapeutic response. For example, several divided doses may beadministered daily with each dose being proportionally reduced.

The compounds of Formulae I-VIII or pharmaceutically acceptable salts ofcompounds of Formulae II-VIII may be administered in a convenientmanner, such as by oral, intravenous (where water soluble), topical,intramuscular or subcutaneous routes or by inhalation.

They may be orally administered, for example, with an inert diluent orwith an assimilable edible carrier, or they may be enclosed in hard orsoft shell gelatin capsules, or compressed into tablets, or incorporateddirectly into the food of the diet. For oral therapeutic administration,they may be incorporated with excipients and used in the form ofingestible tablets, buccal tablets, troches, capsules, elixirs,suspensions, syrups, wafers, and the like. Such compositions andpreparations should contain at least 1% of the compounds of FormulaeI-VIII or pharmaceutically acceptable salts of Formulae II-VIII. Thepercentage of the compositions and preparations may, of course, bevaried and may conveniently be between about 5 to about 80% of theweight of the unit. The amount of the compound of any of Formulae I-VIIIor pharmaceutically acceptable salts of Formulae II-VIII used in suchtherapeutic compositions is such that a suitable dosage will beobtained. Preferred compositions or preparations according to thepresent invention contain between about 25 mg and about 1000 mg of acompounds of Formulae I-VIII or the pharmaceutically acceptable salts ofcompounds of Formulae II-VIII including those containing about 25 mg,about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg, andabout 300 mg.

The tablets, troches, pills, capsules and the like may also contain thefollowing: A binder such as gum tragacanth, acacia, corn starch orgelatin; excipients such as dicalcium phosphate; a disintegrating agentsuch as corn starch, potato starch, alginic acid and the like; alubricant such as magnesium stearate; and a sweetening agent such assucrose, lactose or saccharin may be added or a flavoring agent such aspeppermint, oil of wintergreen, or cherry flavoring. When the dosageunit form is a capsule, it may contain, in addition to materials of theabove type, a liquid carrier.

Various other materials may be present as coatings or otherwise modifythe physical form of the dosage unit. For instance, tablets, pills, orcapsules may be coated with shellac, sugar or both. A syrup or elixirmay contain a compound of Formulae I-VIII or pharmaceutically acceptablesalts of compounds of Formulae II-VIII, sucrose as a sweetening agent,methyl and propylparabens as preservatives, a dye and flavoring such ascherry or orange flavor. Of course, any material used in preparing anydosage unit form should be pharmaceutically pure and substantiallynon-toxic in the amounts employed. In addition, a compound of FormulaeI-VIII or pharmaceutically acceptable salts of compounds of FormulaeII-VIII may be incorporated into sustained-release preparations andformulations. For example, sustained release dosage forms arecontemplated wherein a compound of Formulae I-VIII or pharmaceuticallyacceptable salts of compounds of Formulae II-VIII is bound to an ionexchange resin which, optionally, can be coated with a diffusion barriercoating to modify the release properties of the resin or wherein acompound of Formulae I-VIII or a pharmaceutically acceptable salt ofcompound of Formulae II-VIII is associated with a sustained releasepolymer known in the art, such as hydroxypropylmethylcellulose and thelike.

A compound of Formulae I-VIII or pharmaceutically acceptable salts ofcompounds of Formulae II-VIII may also be administered parenterally. Itis especially advantageous to formulate parenteral compositions indosage unit form for ease of administration and uniformity of dosage.Dispersions can also be prepared in glycerol, liquid polyethyleneglycols, and mixtures thereof and in oils. Under ordinary conditions ofstorage and use, these preparations contain a preservative to preventthe growth of microorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersions. In all cases the form must be sterile and mustbe fluid to the extent that easy syringability exists. It must be stableunder the conditions of manufacture and storage and must be preservedagainst the contaminating action of microorganisms such as bacteria andfungi. The carrier can be a solvent or dispersion medium containing, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyethylene glycol, and the like), suitable mixturesthereof, and vegetable oils. The proper fluidity can be maintained, forexample, by the use of a coating such as lecithin, by the maintenance ofthe required particle size in the case of dispersions and by the use ofsurfactants. The prevention of the action of microorganisms can bebrought about by various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, andthe like. In many cases, it will be preferable to include isotonicagents, for example, sugars or sodium chloride. Prolonged absorption ofthe injectable compositions can be brought about by the use in thecompositions of agents delaying absorption, for example, aluminummonostearate and gelatin.

Sterile injectable solutions are prepared by incorporating a compound ofFormulae I-VIII or pharmaceutically acceptable salts of compounds ofFormulae II-VIII in the required amount in the appropriate solvent withany of the other ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating a sterilized compound of Formulae I-VIII orpharmaceutically acceptable salts of compounds of Formulae II-VIII intoa sterile vehicle which contains the basic dispersion medium and therequired other ingredients from those enumerated above. In the case ofsterile powders, the above solutions are vacuum dried or freeze-dried,as necessary.

A compound of Formulae I-VIII or pharmaceutically acceptable salts ofcompounds of Formulae II-VIII can also be formulated and administered tothe patient in solid or liquid particulate form by directadministration, e.g., inhalation, into the respiratory system.

Solid or liquid particulate forms of a compound of Formulae I-VIII orpharmaceutically acceptable salts of compounds of Formulae II-VIIIprepared for practicing the present invention include particles ofrespirable size: that is, particles of a size sufficiently small to passthrough the mouth and larynx upon inhalation and into the bronchi andalveoli of the lungs. In general, particles ranging from about 1 to 10microns in size are within the respirable range. The pharmaceuticalcompositions containing a compound of Formulae I-VIII orpharmaceutically acceptable salts of compounds of Formulae II-VIII arepreferably administered by direct inhalation into the respiratory systemfor delivery as a mist or other aerosol or dry powder. Particles ofnon-respirable size which are included in the aerosol tend to bedeposited in the throat and swallowed; thus the quantity ofnon-respirable particles in the aerosol is preferably minimized.

In the manufacture of the preferred local formulation, in accordancewith the description herein, a compound of Formulae I-VII orpharmaceutically acceptable salts of compounds of Formulae II-VIII aretypically admixed with, inter alia, an acceptable carrier. The carriermust, of course, be acceptable in the sense of being compatible with anyother ingredients in the formulation and must not be deleterious to thepatient. The carrier may be a solid or a liquid, or both, and ispreferably formulated with the compound as a unit-dose formulation.

Aerosols of liquid particles comprising a compound of Formulae I-VIII orpharmaceutically acceptable salts of compounds of Formulae II-VIII maybe produced by any suitable means, such as inhalatory delivery systems.One is a traditional nebulizer which works in a mechanism similar to thefamiliar perfume atomizer. The airborne particles are generated by a jetof air from either a compressor or compressed gas cylinder-passingthrough the device (pressure driven aerosol nebulizer). In addition,newer forms utilize an ultrasonic nebulizer by vibrating the liquid at aspeed of up to about 1 MHz. See, e.g., U.S. Pat. No. 4,501,729, thecontents of which are incorporated by reference. Nebulizers arecommercially available devices and can transform solutions orsuspensions comprising a compound of Formulae I-VIII or pharmaceuticallyacceptable salts of compounds of Formulae II-VIII into a pharmaceuticalaerosol mist either by means of acceleration of compressed gas,typically air or oxygen, through a narrow venturi orifice or by means ofultrasonic agitation. Suitable formulations for use in nebulizersconsist of a compound of the Formulae I-VIII or pharmaceuticallyacceptable salts of compounds of Formulae II-VIII in a liquid carrier.The carrier is typically water (and most preferably sterile,pyrogen-free water) or a dilute aqueous alcoholic solution, preferablymade isotonic but may be hypertonic with body fluids by the addition of,for example, sodium chloride. Optional additives include preservativesif the formulation is not made sterile, for example, methylhydroxybenzoate, as well as antioxidants, flavoring agents, volatileoils, buffering agents and surfactants, which are normally used in thepreparation of pharmaceutical compositions.

Aerosols of solid particles comprising a compound of Formulae I-VIII orpharmaceutically acceptable salts of compounds of Formulae II-VIII maylikewise be produced with any solid particulate medicament aerosolgenerator. Aerosol generators for administering solid particulatemedicaments to a subject produce particles which are respirable, asexplained above, and generate a volume of aerosol containing apredetermined metered dose of a medicament at a rate suitable for humanadministration. One illustrative type of solid particulate aerosolgenerator is an insufflator. Suitable formulations for administration byinsufflation include finely comminuted powders which may be delivered bymeans of an insufflator or taken into the nasal cavity in the manner ofa snuff. In the insufflator, the powder (e.g., a metered dose thereofeffective to carry out the treatments described herein) is contained incapsules or cartridges, typically made of gelatin or plastic, which areeither pierced or opened in situ and the powder delivered by air drawnthrough the device upon inhalation or by means of a manually-operatedpump. The powder employed in the insufflator consists either solely ofthe compound of Formulae I-VIII or pharmaceutically acceptable salts ofcompounds of Formulae II-VIII or of a powder blend comprising a compoundof the Formulae I-VIII or pharmaceutically acceptable salts of compoundsof Formulae II-VIII a suitable powder diluent, such as lactose, and anoptional surfactant. A second type of illustrative aerosol generatorcomprises a metered dose inhaler. Metered dose inhalers are pressurizedaerosol dispensers, typically containing a suspension or solutionformulation of a compound of Formulae I-VIII in a liquefied propellant.During use, these devices discharge the formulation through a valve,adapted to deliver a metered volume, from about 10 to about 22microliters to produce a fine particle spray containing tetracycline.

Any propellant may be used in carrying out the present invention,including both chlorofluorocarbon-containing propellants andnon-chlorofluorocarbon-containing propellants. Fluorocarbon aerosolpropellants that may be employed in carrying out the present inventionincluding fluorocarbon propellants in which all hydrogens are replacedwith fluorine, chlorofluorocarbon propellants in which all hydrogens arereplaced with chlorine and at least one fluorine, hydrogen-containingfluorocarbon propellants, and hydrogen-containing chlorofluorocarbonpropellants. Examples of such propellants include, but are not limitedto: CF₃CHFCF₂, CF₃CH₂CF₂H, CF₃CHFCF₃, CF₃CH₂CF₃) CF₃CHCl—CF₂Cl,CF₃CHCl—CF₃, CF₃CHCl—CH₂Cl, CF₃CHF—CF₂Cl, and the like. A stabilizersuch as a fluoropolymer may optionally be included in formulations offluorocarbon propellants, such as described in U.S. Pat. No. 5,376,359to Johnson. The aerosol formulation may additionally contain one or moreco-solvents, for example, ethanol, surfactants, such as oleic acid orsorbitan trioleate, antioxidants and suitable flavoring agents.

Compositions containing respirable dry particles of micronized compoundsof any of Formulae I-VIII or pharmaceutically acceptable salts ofcompounds of Formulae D-VIII may be prepared by grinding the dry activecompound, with e.g., a mortar and pestle or other appropriate grindingdevice, and then passing the micronized composition through a 400 meshscreen to break up or separate out large agglomerates.

The aerosol, whether formed from solid or liquid particles, may beproduced by the aerosol generator at a rate of from about 10 to 150liters per minute. Aerosols containing greater amounts of a compound ofFormulae I-VIII or pharmaceutically acceptable salts of compounds ofFormulae II-VIII may be administered more rapidly. Typically, eachaerosol may be delivered to the patient for a period from about 30seconds to about 20 minutes, with a delivery period of about 1 to 5minutes being preferred.

The particulate composition comprising a compound of Formulae I-VIII orpharmaceutically acceptable salts of compounds of Formulae II-VIII mayoptionally contain a carrier which serves to facilitate the formation ofan aerosol. A suitable carrier is lactose, which may be blended with thetetracycline in any suitable ratio.

The compounds of Formulae I-VIII or pharmaceutically acceptable salts ofcompounds of Formulae II-VIII may be formulated into a topicalcomposition. Typically, such topical compositions are prepared as aliquid (both oil-in-water and water-in-oil, with or without emulsions),gels (both aqueous and non-aqueous), lotions, ointments, paste, powdersand the like. The compounds of Formulae I-VIII or pharmaceuticallyacceptable salts of compounds of Formulae II-VIII are present intherapeutically effective amounts as defined herein. Preferably, thetopical composition of the present invention contain from about 0.5% toabout 95% by weight of compounds of Formulae I-VIII or pharmaceuticallyacceptable salts of compounds of Formulae II-VIII.

The topical compositions are most useful in treating dermatologicdisorders or conditions arising from or caused by excess IgE in theplasma in a mammal, such as a topic dermatosis, inflammatory dermatosis,such as acne, and the like.

Cream, lotion, gel, stick, ointment, or topical solution, may be used astopical vehicles for compounds of the formula (I) in conventionalformulations well known in the art, for example, as described in thestandard text books of pharmaceutics and cosmetics, such as Harry'sCosmeticology published by Leonard Hill Books, Remington'sPharmaceutical Sciences published by Mack Publishing Co., and theBritish and US Pharmacopoeias. A standard emulsifying ointment base orglycerol or anhydrous polyethylene glycol are simple examples ofsuitable vehicles.

Examples of oils suitable for inclusion in a standard emulsifyingointment base include mineral oils, vegetable oils, synthetic fatty acidesters, fatty-alcohols, lanolin and its derivatives.

These compositions will normally include a suitable emulsifier. Thecomposition can range from liquid through semi-liquid to gel typesaccording to the type of emulsion and quantity of any thickening agentwhich may be present. Examples of emulsifiers include polyhydric alcoholesters such as sorbitan monostearate, fatty acid esters such as glycerylmonostearate, and polyester derivatives of fatty acids or fattyalcohols.

These topical compositions as well as the other pharmaceuticalcompositions described hereinabove may also contain anti-oxidants andother conventional ingredients such as preservatives, humectants,perfumes and alcohol. Advantageously, a penetrating agent such as AZONEmay also be included.

The compositions for topical treatment may also contain othertherapeutic agents such as anti-infective and/or anti-viral agents.Suitable anti-infective agents include the topically applicableantibacterial, anti-yeast, anti-fungal and anti-herpes agents.

The topical composition of the present invention can also be applied intherapeutic effective amounts through a transdermal patch usingtechniques known to one of ordinary skill in the art.

The compounds of any of Formulae I-VIII or pharmaceutically acceptablesalts of compounds of Formulae II-VIE are administered by these variousmodes of administration in therapeutically effective amounts, as definedherein which amounts are determined by the physician.

The pharmaceutical compositions of the present invention are inassociation with a pharmaceutically acceptable carrier, and contain theingredients, typically found in pharmaceutical compositions. Forexample, it may include excipients, fragrances, anti-oxidants,sunscreens, flavors, buffers, solubilizers, thickeners and the like. Asused herein, “pharmaceutically acceptable carrier” includes any and allsolvents, dispersion media, coatings, antibacterial and antifungalagents, isotonic and absorption delaying agents for pharmaceuticalactive substances well known in the art. Except insofar as anyconventional media or agent is incompatible with the compounds ofFormulae I-VIII or pharmaceutically acceptable salts of compounds ofFormulae II-VIII their use in the therapeutic compositions iscontemplated. Supplementary active ingredients can also be incorporatedinto the compositions.

Dosage unit form as used herein refers to physically discrete unitssuited as unitary dosages for the subjects to be treated; each unitcontaining a predetermined quantity of active material calculated toproduce the desired therapeutic effect in association with the requiredpharmaceutical carrier.

The compounds of Formulae I-VIII or pharmaceutically acceptable salts ofcompounds of Formulae II-VIII are compounded for convenient andeffective administration in effective amounts with a suitablepharmaceutically acceptable carrier in dosage unit form as hereinbeforedescribed. A unit dosage, for example, contains a compound of FormulaeI-VIII or pharmaceutically acceptable salts of compounds of FormulaeII-VIII singly or in total in amounts ranging from about 50 mg to about1000 mg. If placed in solution, the concentration of the compounds ofFormulae I-VIII or pharmaceutically acceptable salts of compounds ofFormulae II-VIII singly or in combination preferably ranges from about10 mg/ml to about 250 mg/ml. The preferred mode of administration isoral.

Moreover in normal humans, not suffering from asthma or allergies ordiseases wherein IgE is pathogenic, the administration of compounds ofFormulae I-VIII or pharmaceutically acceptable salts of compounds ofFormulae II-VIII does not increase the concentration of IgE in theplasma.

The compounds of each of Formulae I-VIII or pharmaceutically acceptablesalts of compounds of Formulae II-VIII are useful in treating asthma.They are anti-inflammatory agents and can be used to treat inflammatoryconditions.

The present inventors have found that the higher the concentration ofthe free IgE in the plasma in those patients suffering from a diseasewhere IgE is pathogenic, the greater is the risk of the disease statebecoming aggravated or worsening. By lowering the amount of the free IgEin the plasma of those patients, the physician will lower the risk ofthe disease state becoming worse or becoming exacerbated.

In another embodiment, the present invention is directed to theprophylaxis of the disease wherein IgE is pathogenic from becoming moresevere, which method comprises administering to the patient aprophylactically effective amount of any of the compounds of FormulaeI-VII, or pharmaceutically acceptable salts of compounds of FormulaeII-VIII individually or combination thereof. The effective amount inthis embodiment is determined by the physician; usually, this amount isthe same as the therapeutic effective amount, that was describedhereinabove.

A compound of Formulae I-VIII or pharmaceutically acceptable salts ofcompounds of Formulae II-VIII can also be given to mammals sufferingfrom asthma, or allergies or other diseases where IgE is pathogenic byadministering to the patient a prophylactically effective amount of acompound of Formulae I-VIII or combination thereof. The effectiveamounts can be determined by the physician. The normal person has a freeplasma concentration of IgE of less than about 50 IU/ml. Thus, theadministration of the aforementioned compounds of Formulae I-VIII orpharmaceutically acceptable salts of compounds of Formulae II-VIII willprevent and/or retard the onset of allergy or allergic asthma and/orother disease where IgE is pathogenic when administered in effectiveamounts, as described herein. In addition, the administration thereofwill prevent or lower the risk of persons having a free IgEconcentration in the plasma increasing to the level where the person isat a substantial risk of suffering from a disease where IgE ispathogenic. Preferably, the free IgE level in the plasma, after suchtreatment, will remain less than about 50 IU/ml.

In the present specification, unless indicated to the contrary, theplural will connote the singular and vice versa.

The term “treating”, “treat” or “treatment” used herein refers to thereduction and/or alleviation of at least one adverse effect or symptomof a disease where IgE is pathogenic. It refers to the management andcare of a mammalian subject, preferably humans, for the purpose ofcombating the disease, conditions or disorders where IgE is pathogenic,and includes the administration of any of the compounds of FormulaeI-VIII, alone or in combination to delay the onset of at least onesymptom or complication associated with the disease, alleviating thesymptom or effect or complications associated therewith or in thealternative eliminating the disease or condition.

The term “prophylaxis” or “prevent” or synonym thereto refers to theprevention or a measurable reduction in the likelihood of a patientacquiring a disease where IgE is pathogenic, even if the mammal issuffering form another malady which debilitates it and makes it moresusceptible to such a disease. If a patient or mammal is suffering froma disease where IgE is pathogenic, the term also refers to the reductionin the likelihood of the disease becoming acerbated.

The term “therapeutically effective amount” is synonymous with “IgElowering effective amounts” and refers to the amount effective ineliminating or alleviating or curing the symptoms associated with adisease or malady where IgE is pathogenic and/or alleviating or curingthe disease altogether.

The term “prophylactically effective amount” refers to the amounteffective in preventing or reducing the likelihood of a mammal, e.g.,patient from acquiring a disease in which IgE is pathogenic. It alsorefers to the amount effective in preventing a mammal afflicted with adisease or malady where IgE is pathogenic from worsening or becomingmore severe.

As indicated herein, the prophylactive effective amounts and thetherapeutically effective amounts can be determined by the physician;however, it is preferred that these amounts are the same.

As used herein, the term compounds of Formulae I-VIII refers to a singlecompound or combination of compounds having Formulae I-VIII.

As used herein, the singular denotes the plural and vice versa.

The compounds of the present invention can be prepared by art recognizedtechniques as shown hereinbelow.

Compounds of Formula I can be prepared as follows as shown in Scheme I:

For example, the phenol derivative (IA) is reacted with acetyl chloridein the presence of strong base, such as sodium amide, sodium hydride andthe like in an inert solvent, such as THF under effective reactedconditions to form a compound of Formula IB. It, in turn, is reactedwith acetyl chloride in the presence of a stronger base such asLi(N-i-Pr)₂, in an inert solvent, such as THF to form Compound IC.Compound IC enolizes to form a compound a Formula I.

Compounds of Formula II can be prepared as follows:

For example, the compound of Formula IIA is reacted with a diene ofFormula IIB under Diels Alder reaction conditions. Preferably, thereaction is conducted in the presence of a Lewis acid catalyst, e.g.,zinc halide, such as ZnI₂, and in an inert solvent, such as toluene,xylene, benzene and the like, and preferably xylene, under conditionssufficient to form the six membered cyclic diene EC. The reaction may beconducted under reflux, although temperatures from about 20° C. toreflux may be used. As used hereinabove, TBDMS is t-butyl dimethylsilyl,Z is a hydroxy protecting group known in the art, and P is a chiralgroup, such as (+) or (−) phenylmethyl. Various hydroxy protectinggroups can be utilized, such as benzyl, carbonate and the like, andthese are described on pages 10-72 of the book entitled ProtectiveGroups in Organic Synthesis, by Theodora W. Greene, John Wiley & Sons,New York, N.Y., 1981, the contents of which are incorporated byreference. The cyclohexane IIC is oxidized with an oxidizing agent toform an epoxide. Preferably, the cyclohexane is reacted withm-chloroperbenzoic acid, H₂O₂ and the like under epoxidizing conditionssufficient to form an epoxide alcohol of Formula IID. The alcohol HD isoxidized with an oxidizing agent to form the corresponding ketone HE.Oxidizing groups known in the art, such as CrO₃ or K₂Cr₂O₇ and the like,may be utilized as the oxidizing agent. In addition, the alcohol HD canbe oxidized to HE using oxalyl dimethyl sulfoxide in combination withone of the following reagents: acetic anhydride,SO₃-pyridine-triethylamine, trifluoroacetic anhydride, oxalyl chloride,tosyl chloride, chlorine, bromine, AgB F₄-Et₃N, P₂O₅-Et₃N, phenyldichlorophosphate, trichloromethyl chloroformate, trimethylamine oxide,and the like. It is preferred that the reaction takes place using DMSOand oxalyl chloride in the presence of an amine base under Swernoxidation conditions. The epoxide HE is oxidized to the correspondingalpha-hydroxy ketone IIF with conventional oxidizing agents such as Br₂.N-bromosuccinimide and the like. The Compound IIF is in equilibrium withthe corresponding enol. The ester moiety of Formula IIF is reacted withammonia to form the corresponding amide under effective amide formingconditions to form the compound of Formula II.

An exemplary procedure for preparing the compound of Formula III is asfollows:

IIIA is reacted with a HCOCOOnBu and base, such as sodium hydride orsodium amide, under Claisen Condensation Reaction conditions (i.e.,α-arylalkyl dialkoxy substitution reaction conditions) to form IIIB,which in turn is reacted with dimethylamine under nucleophilic reactionconditions to form IIIC. IIIC can be further converted to IIID, usingthe procedures described in Woodward, et al, JACS 84, 3222-3224, (1962)the contents of which are incorporated by reference. IIID is reactedwith 0% in DMF and MeOH at pH 5 to afford III.

An exemplary procedure for the preparation of Formula IV is as follows:

This is prepared in an analogues manner as described for preparing thecompound of Formula III. The diketone, represented as its enol-ketoneIVA, is reacted with HCOCOOn-Bu under Claisen Condensation conditions,(i.e., α-acylalkyldealkoxy substitution reaction conditions) to formIVB, which is reacted with dimethylamine under nucleophilic reactionconditions to form IVC. IVC is converted to IVD using the proceduresdescribed by Woodward, et al. in JACS, 84, 3222-3224 (1962), thecontents of which are incorporated by reference to give IVD. IVD isreacted with O₂ in DMF and MeOH at pH 5 to afford IV.

An exemplary procedure of the preparation of the compound of Formula Vis as follows:

One of the ketone moieties in VA is protected with protection groupsknown in the art, such as by reacting with ethylene glycol, MeOH in acidand the like. The non-protected ketone is reacted with an organometalliccompound such as CH₃M, wherein M is a metal, such as lithium, e.g.,LiCH₃ or Grignard reagent, such as CH₃MgX, wherein X is halide, whichproduct is reduced in the presence of a reducing agent known in the art,boranes to reduce the resulting alcohol. The protecting group is removedto afford the ketone. VB is reacted with a succinic acid derivative,e.g., ester, such as C₁-C₉ alkyl succinate, e.g., such as methylsuccinate, in the presence of base under Michael-like addition reactionconditions to form the product VC which, in the presence of base andheat decarboxylates to produce VD. VD is esterified with alcohol such asMeOH in acid and the product thereof is reacted with methyl carbonate inbase to form the product VE in situ under Claisen-like condensationreaction conditions, which in base, such as sodium hydride cyclizesunder Dieckman condensation reaction conditions to form the product VF.This is in turn decarboxylates in acid and heat to form VG. In thepresence of HCO COOnBu and base under Claisen condensation reactionconditions, product VH is formed, as in the preparation of Compound III.This in turn is reacted with dimethylamine under nucleophilic conditionsto form the product VJ. VJ in the presence of base, such as sodiumhydride and heat, such as 120° C., in DMF in accordance with theprocedure described in Woodward, et al. JACS 84, 3222-3224 (1962), thecontents of which are incorporated by reference, cyclizes to afford VK.VK is heated in xylene at a temperature sufficient to eliminatecyclopentadiene, such as from about 120° C. to 160° C., and theresulting intermediate is further reacted with O₂ in DMF and MeOH at pH5 to afford VL which is reduced with a reducing agent using techniquesknown in the art, such as by hydrogenation to form V.

Compounds of Formula VI can be prepared as follows:

One of the ketone moieties in VIA is protected with protection groupsknown in the art, such as by reacting it with ethylene glycol, MeOH inacid and the like. The mono-protected ketone is reacted with a reducingagent known in the art to reduce ketones to alcohols and then alkanes,such as NaBH—AlCls, borane, and the like. The protecting group isremoved to afford the ketone VIB. VIB is reacted with a succinic acidderivative, such as methyl succinate, in the presence of base underMichael-like addition reaction conditions to form the product VIC, whichin the presence of base and heat decarboxylates to produce VID. VID isesterified with alcohol, such as MeOH in acid and the product thereof isreacted with methyl oxalate in base to form the product VIE in situunder Claisen-like condensation reaction conditions, which in base suchas NaH, cyclizes under Dieckman condensation reaction conditions to formthe product VIF. This is in turn decarboxylates in acid and heat to formVIG. In the presence of HCO—COOnBu and base under Claisen condensationreaction as described in the preparation of compound III, product VIH isformed. This in turn is reacted with dimethylamine under nucleophilicconditions to form the product VIJ, VIJ is converted to VIK, using theprocedures described in Woodward, et al. JACS 84, 3222-3224 (1962), thecontents of which are incorporated by reference. VIK is heated in xyleneunder conditions sufficient to eliminate cyclopentadiene, such as fromabout 120° C. to about 160° C., and the resulting intermediate isreacted with 0% in DMF and MeOH at pH 5 to afford VIL. A carbon-carbondouble is reduced using a reducing agent by techniques known in the art,such as hydrogenation, to form VI.

Compound VII can be prepared as shown in the following scheme:

Compound VII is prepared as follows, 2-cyclohexen-1-one is reacted withethylene glycol in the presence of acid, such as putoluenesulfonic acid(TSOH) under conditions to form a Ketal protected cyclohexene (VIIB) asdescribed in the article by Laronte, et al. in Synthetic Communications.21(7), 881-884 (1991). It is to be noted that protection of VIIA wasaccompanied by deconjunction of the double bond (isomerism) to formVIIB. Oxidation of VIIB using strong oxidizing agents, such as peracids,e.g., meta-chloro perbenzoic acid (MCPBA) produced the epoxide (VIIC).Reaction of the epoxide with dimethylamine in mineral acid, such ashydrochloric acids, sulfuric acid and the like, produced the aminoalcohols VIID and VIIE. Separation of VIID from VIIE is effected usingseparation techniques known in the art, such as chromatography, e.g.,HPLC chromatography, column chromatography and the like. VIID isoxidized by an oxidizing agent known in art such as acid dichromate,KMnO₄, Br₂, MnO₂, ruthenium tetroxide, chromic acid and sulfuric acid inwater, PyrSO₃, wherein Pyr is pyridine, and the like, to form VHF.Removal of the ketal protecting groups affords VIIG, which is inequilibrium with the enol form VII.

Compound VIII can be prepared by art recognized techniques as follows:

Protection of 1,4-cyclohexadione (VIIIA) with, ethylene glycol providesthe protected keto ketal (VIIIB). The ketal VIIIB is reacted with methylcarbonate and strong base, such as potassium hydride under Claisencondensation reaction conditions, such as in the preparation of compoundIII, described hereinabove (See Synthesis. 1249-54, (1994). The productthereof is reduced with a reducing agent, such as sodium borohydride,H₂/Pt and the like, (See Synlett. 4, 497-500 (2000)) and the resultingproduct is reacted with triisopropylsilicon chloride in the presence ofbase, such as diisopropylethylamine (Hunigs base) to provide a compoundof formula VIIIC. Compound VIIIC is reacted with, acid, such asp-toluenesulfonic acid, to afford the product VIIID, where TIPS istriisopropylsilyl (See, e.g., Journal of Org. Chem., 66(11) 3653-3661(2001), the contents of which are incorporated by reference). VIIID isreacted with dimethylamine in the presence of a reducing agent whichreduces the resulting imine to produce the amine of Formula VIIIE. Theamine is reacted with tetrabutylammonium fluoride followed by oxidationwith PhSeCl and peroxide to form the product of Formula VIIIF (See,Synthesis 9, 850-853 (1987) and Tetrahedron Lett. 32(42), 5953-5956(1991)). The ester is hydrolyzed with acid and then reacted with NH₃under amide forming conditions to afford the product VIII.

Alternatively, the compound of the Formula VIII is formed as follows:

IXA is subjected to conditions to form the nitronium, IXB. Morespecifically, IXA is reacted with, a weak organic base, such as tertiaryamine, e.g., a salt of diisopropylethylamine salt, in the presence of aninert solvent, such as methylene chloride under conditions effective toform the aziridinium ion, e.g., at temperatures ranging from about 0° C.to the reflux temperature of the solvent, but preferably at about 0° C.The aziridinium ion IXB, which is formed in-situ is reacted with IX inthe presence of a weak base such as a tertiary amine under conditionssufficient to form IXC. IXC is converted to IXD as described in SchemeVIII by reacting IXC with a reducing agent such as Na BH₄ and then withTIPSCl followed by reacting the product thus formed with an acid such asTsOH; the product thereof is reacted with tetrabutylammonium fluoridefollowed by oxidation with PhSeCl and peroxide to form the product 1×D.The ester in DOD is hydrolyzed in acid and then reacted with NH₃ underamide forming conditions to afford the product VIII: which is convertedto VIII as described in Scheme VIII.

The compounds of the present invention are effective in lowering theexcess IgE concentrations in the plasma. However, they do not sufferfrom the disadvantages described hereinabove associated with thetetracyclines, minocycline or doxycyclines. Moreover, since thecompounds of Formulae I-VIII are smaller molecules, they have a higherbioavailability than the corresponding tetracyclines and thus higheramounts of these molecules can be absorbed into the blood and plasmathan the corresponding tetracyclines. Thus, significantly less drug isrequired to be administered relative to the tetracyclines for efficacy.

While the foregoing specification teaches the principle of the presentinvention, with examples provided for the purposes of illustration,these teachings will make apparent to those skilled in the art otherembodiments and examples. These other embodiments and examples are alsowithin the scope of the present invention.

1. (canceled)
 2. A compound or pharmaceutically acceptable salt thereofwhere the compound has the formula


3. A compound or pharmaceutically acceptable salt thereof where thecompound has the formula


4. A compound or pharmaceutically acceptable salt thereof where thecompound has the formula


5. A compound or pharmaceutically acceptable salt thereof where thecompound has the formula


6. A compound or pharmaceutically acceptable salt thereof where thecompound has the formula


7. A compound or pharmaceutically acceptable salt thereof where thecompound has the formula


8. A compound or pharmaceutically acceptable salt thereof where thecompound has the formula

9.-22. (canceled)